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目的比较浒苔(Enteromorpha)多糖4种组分EP,EP1、EP2和EP3对体外培养的小鼠T、B淋巴细胞增殖的影响,并观察作用效果最佳组分EP2在T、B淋巴细胞中的分布及其对细胞RNA合成的影响。方法采用Sevage法,Sephacryl-300分子筛层析以及乙醇分级沉淀法分离纯化获得4个浒苔多糖组分EP,EP1、EP2和EP3,通过体外细胞增殖实验评价不同组分的免疫学活性,另外,利用PAS染色法和甲基绿-派洛宁染色法显示细胞内多糖分布及RNA合成情况。结果 EP、EP1、EP2和EP3中单糖、糖醛酸、硫酸根含量存在差异,均能促进B淋巴细胞以及ConA或LPS诱导的T、B淋巴细胞增殖,且EP2的作用效果最好;经EP2处理的T、B淋巴细胞或同时用EP2和丝裂原(ConA或LPS)处理的T、B淋巴细胞体积明显增大,RNA合成增加,B淋巴细胞有母细胞化趋势。结论浒苔多糖能调节小鼠淋巴细胞活性,促进丝裂原诱导的淋巴细胞分化和免疫应答,有望成为1种新的动物免疫增强剂。
Objective To compare the effects of Enteromorpha polysaccharides (EPom), EP1, EP2 and EP3 on the proliferation of T and B lymphocytes in vitro and to observe the effect of EP2 in T and B lymphocytes Distribution and Its Effect on Cell RNA Synthesis. METHODS: Sephacryl-300, Sephacryl-300 and ethanol fractionation methods were used to isolate and purify 4 Enteromorpha polysaccharides components EP, EP1, EP2 and EP3. The immunological activities of different components were evaluated by in vitro cell proliferation assay. The distribution of intracellular polysaccharides and the synthesis of RNA were revealed by PAS staining and Methyl Green - Peyronine staining. Results The differences of monosaccharide, uronic acid and sulfate in EP, EP1, EP2 and EP3 all promoted the proliferation of B lymphocytes and T and B lymphocytes induced by ConA or LPS, and the effect of EP2 was the best. The volume of T and B lymphocytes treated with EP2-treated T, B lymphocytes or both EP2 and mitogen (ConA or LPS) increased significantly, the RNA synthesis increased, and B lymphocytes had the tendency of baryocytosis. CONCLUSION: Enteromorpha prolifera polysaccharide can modulate lymphocyte activity of mouse and promote mitogen-induced lymphocyte differentiation and immune response, which is expected to become a new animal immune enhancer.